摘要
目的:研究雷公藤内酯醇(triptolid,TPL)与热疗联合应用对胃癌细胞MKN28增殖、细胞周期分布及磷酸化蛋白激酶B(phosphorylation of Akt,p-Akt)表达的影响。方法:将体外复苏和培养的胃癌细胞系MKN28分为四个组:对照组、TPL组、热疗组、TPL联合热疗组。其中TPL刺激浓度为20ng/ml,热疗采用43℃下加热150min。使用四甲基偶氮唑盐比色法(measurement of trititaed thymidine incorporation,MTT)检测细胞增殖并计算细胞增殖抑制率。流式细胞仪(flow cytometer,FCM)检测细胞周期分布的变化,使用蛋白质印迹法(Western blot)检测p-Akt水平。结果:热疗组,TPL组和TPL联合热疗组MKN28细胞增殖抑制率分别为21.2%,28.5%和54.5%,联合组显著高于TPL组和热疗组。FCM结果显示热疗组G0/G1期减少至(62.7±3.8)%,G2/M期增多至(18.1±3.2)%。TPL组G0/G1期增多至(80.62±4.6)%,G2/M期增多至(16.18±2.2)%。联合组G0/G1期(77.72±8.1)%,G2/M期增多至(19.62±2.4)%。热疗组和TPL组对细胞周期的影响不一样。与热疗组和TPL组相比,联合组G2/M期增多最明显。Western blot检测各组p-Akt蛋白表达均有降低,其中对照组(1.81±0.05),热疗组(1.28±0.01),TPL组(0.95±0.07),联合组(0.58±0.06),联合组下降最明显(P<0.05)。结论:TPL联合热疗对MKN28细胞增殖抑制作用更强,可能与二者对细胞周期的作用点不同,并能同时降低细胞内p-Akt的表达有关。
Objective:To explore the cytotoxic effects of TPL and hyperthermia in human gastric cancer cells in vitro.Methods:MKN28 cells were treated by four methods:control group,TPL group,hyperthermia group and TPL combined hyperthermia group.MTT assay was adopted to investigate the proliferation rates of cells.FCM was used to check the change of cell cycle in vitro.P-Akt of each group was checked by Western blot.Results:TPL combined hyperthermia group could significantly inhibit cell proliferation as compared with TPL group or hyperthermia group alone.The outcome of FCM indicated that TPL and hyperthermia affected cell-cycle distribution.The number of cells in G2/M phase had the greatest increase in TPL combined hyperthermia group.The level of p-Akt protein was lower in TPL group(0.95±0.07),hyperthermia group(1.28±0.01) and combined group(0.58±0.06) when compared with those in the control group (1.81±0.05),and the differences were significant(P〈0.05).Conclusion:TPL with thermotherapy can enhance therapeutic effect on gastric tumor cells.It may be related with the distribution of cell cycle and the inhibition of expression of p-Akt.
出处
《现代肿瘤医学》
CAS
2014年第1期38-40,共3页
Journal of Modern Oncology
基金
湖北省卫生厅青年科技人才项目(No.QJX2012-08)
中央财政专项经费(No.2012QN050)